Glucose oxidase (GOD) is an important industrial enzyme with many potential applications. In order to increase the production and productivity of GOD by recombinant Pichia pastoris GS115, we investigated the feeding strategies of mixed carbon sources during induction phase, based on results of the optimization of initial cell and methanol concentration on GOD production. The optimal initial cell and methanol concentration were 100 g/L and 18 g/L. During induction phase, the mixed-carbon-sources strategies showed that glycerol, sorbitol or mannitol co-feeding with methanol could enhance GOD production. With mannitol co-feeding (20:1(W/W)), the maximum GOD production and maximum GOD productivity reached 711.3 U/mL and 4.60 U/(mL x h) after an induction period of 156 h. Compared to the control, the enhancements of GOD production and productivity were 66.3% and 67.9%, respectively. Meanwhile, we found an appropriate mannitol co-feeding strategy that would not inhibit the expression of promote. The activity of alcohol oxidase was 8.8 U/g, which was enhanced by 69.2% compared to the control (5.2 U/g). We can use the same optimization process to improve the production of other proteins from recombinant Pichia pastoris by changing the fermentation parameters.
Carbon ; metabolism ; Fermentation ; Glucose Oxidase ; biosynthesis ; Glycerol ; metabolism ; Industrial Microbiology ; Mannitol ; metabolism ; Methanol ; metabolism ; Pichia ; metabolism ; Sorbitol ; metabolism

Objective To cross-cultural adapt and test the reliability and validity of Humanistic Relationship Experience Scale (HRES) made by Boscart. Methods Cross-culture Adaptation was carried out by following American Academy of Orthopaedic Surgeons Evidence Based Medicine Committee (AAOS). A convenience sampling was conducted among 180 patients with chronic disease, and then assessed the reliability and validity of HRES. The internal consistency coefficient and the test-retest were used to assess the reliability of HRES, and the content validity index (CVI) and construct validity were used to assess the validity of HRES. Results In order to understand the purpose and content of the scale easily, the Chinese version title of HRES was revised to"Nursing Humanistic Care Experience Scale-Noninfectious Chronic Disease Patients Version (NHCES-NCDP)" after soliciting experts′ advice. The CVI of the scale was 1;factor analysis got three factors, which could explain 67.897％of the total variance. A 0.965 Cronbachαof total scale was obtained, and for each dimension range was 0.876-0.948;the test-retest reliability was 0.710 for the overall scale. Conclusion NHCES-NCDP is reliable and valid, which can be used to assess the experience of nursing humanistic care for patients with chronic diseases, and to explore nurses′implementation of nursing humanistic care.

Objective:This study is to investigate the predictive value of serum levels of TIMP-2 and insulin-like growth factor-binding protein 7(IGFBP7) in patients with DCD(donation after cardiac death) kidney transplantation.Methods:A prospective research design was used to select DCD kidney transplant patients admitted to the Li Huili Hospital of Ningbo University from January 2018 to October 2020.Inclusion criteria: ①Complete data; ②There were no serious complications affecting the function of the transplanted kidney in the early postoperative period.Exclusion criteria: ①Incomplete data; ②Patients were unable or unwilling to cooperate with the study; ③Severe complications affecting the function of the transplanted kidney occurred early after the operation.The ELASE method was used to quantitatively detect the serum TIMP-2 and IGFBP7 levels at 6, 12, 24, 48, 72 hours and 7 days after renal transplantation, and monitor the serum creatinine values during the same period and 21 days after the operation. According to the occurrence of DGF, the measured values of TIMP-2 and IGFBP7 at different time points and their product's ability to predict the occurrence of DGF after kidney transplantation were analyzed. The receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to evaluate the diagnostic efficacy of TIMP-2 and IGFBP7 for DGF.Results:A total of 33 patients were enrolled, 7 patients (21.2%) in the DGF group and 26 patients (78.8%) in the non-DGF group. Between the two groups, the donor glomerular filtration rate were [98.5(15.8-132.5)ml/(min·1.73m 2) and 79.1(60.6-102.5)ml/(min·1.73m 2)], recipient gender (male/female: 3/4 cases and 10/16 cases), recipient age [48(34-56) Years old and 45(23-61) years old], the recipient's preoperative creatinine [1114.0(731.4-1293.0)μmol/L and 858.4(657.6-1051.9)μmol/L], the recipient's preoperative urea nitrogen [15.0(13.2-19.6)mmol/L and 17.3(13.6-20.9)mmol/L], receptor preoperative albumin [43.5(38.5-45.3)mmol/L and 41.2(37.5-46.1) mmol/L], recipient dialysis method [hemodialysis/peritoneal dialysis: 3/4 cases and 9/17 cases], warm ischemia time [6(5-7) and 5(4-6) min, there was no statistically significant difference] ( P>0.05). The values of serum IGFBP7 and TIMP-2×IGFBP7 in the DGF group were higher than those in the non-DGF group at all time points ( F=15.753, P=0.040; F=13.000, P=0.024), while serum TIMP-2 was not significant between the two groups difference ( F=1.157, P=0.075). For the diagnostic value of DGF, the AUC of serum IGFBP7 at 48 h after surgery was 0.863 (95% CI 0.696-1.000, P=0.004). When 5.97 ng/ml was used as the cut-off value, the sensitivity was 85.7% and the specificity was 80.8 %. The AUC of TIMP-2×IGFBP7 at 48 hours after surgery was 0.819 (95% CI 0.641-0.996, P=0.011). When 62.06(ng/ml) 2 was used as the cutoff value, the sensitivity was 71.4% and the specificity was 80.8%.There was no statistical difference in the area under the curve between the two ( P>0.05). There were differences in the dynamic trend of serum IGFBP7 and creatinine in the DGF group. Serum IGFBP7 at 7 days after surgery was positively correlated with creatinine at 21 days after surgery. Conclusion:Serum IGFBP7 and TIMP-2×IGFBP7 could predict the occurrence of DGF after DCD donor kidney surgery. The predictive value changes with time. Among them, 48h and 7d after surgery are the most valuable. However, serum TIMP-2 has not been found to have predictive value in this study.

Objective:To explore the predictive values of tissue inhibitor of metalloproteinase-2 (TIMP-2) and insulin-like growth factor binding protein 7 (IGFBP7) in donor sera and lavage fluid on delayed graft function (DGF) in donation after circulatory death (DCD) kidney transplant recipients.Methods:A total of 33 eligible kidney donors and 33 corresponding recipients were recruited. Preoperative serum and renal perfusion fluid samples of donors were collected to determine the levels of TIMP-2 and IGFBP7. Patients were grouped according to whether DGF occurred after kidney transplantation and measured indicators analyzed. Independent sample t test was utilized for comparing the groups with normal distribution measurement data. And χ2 test was employed for comparing the groups with normal distribution counting data and Mann-Whitney test for comparing the groups with non-normal distribution measurement data. Receiver operating characteristic (ROC) curve and area under curve (AUC) were used for evaluating the diagnostic efficacy of indicators. Results:In donor-DGF group, lavage fluid TIMP-2, product of lavage fluid TIMP-2 and IGFBP7 (TIMP-2×IGFBP7), serum IGFBP7 and product of serum TIMP-2 and IGFBP7 (TIMP-2×IGFBP7) were higher than those in donor-non-DGF group ( P<0.05). The AUC of TIMP-2, TIMP-2×IGFBP7, serum IGFBP7 and serum TIMP-2×IGFBP7 in the diagnosis of DGF were 0.753 (95%CI 0.546~0.959), 0.747 (95%CI 0.510~0.984), 0.824 (95%CI 0.615~1.000) and 0.852 (95%CI 0.660~1.000) respectively. Conclusions:Donor serum IGFBP7, donor serum TIMP-2×IGFBP7, lavage fluid TIMP-2 and lavage fluid TIMP-2×IGFBP7 may be used for predicting the occurrence of early DGF after kidney transplantation. Among them, serum TIMP-2×IGFBP7 has the highest diagnostic efficiency and may be an excellent predictor of DGF occurrence.

Objective:To analyze the incidence of liver function injury in patients infected with 2019-nCoV omicron variant and its influencing factors.Methods:The clinical data and laboratory findings of 897 COVID-19 patients infected with omicron variant in Zhejiang province from February 23 to July 14, 2022 were retrospectively analyzed. Patients were divide into liver function injury group ( n=243) and non-liver function injury group ( n=654) based on liver function indicators. The clinical characteristics and laboratory tests were compared between the two groups, and influencing factors of liver function injury were analyzed. SPSS 26.0 statistical software was used for data analysis. Results:The incidence of liver injury in this series was 27.09% (243/897). The median age of patients in liver injury group was older, the body mass index (BMI) was higher( Z=-6.237 and -2.166, both P<0.05), the proportions of patients with hypertension and diabetes, and with severe clinical classification were higher ( χ2=17.087, 27.509 and 12.945, all P<0.01) ; the proportion of vaccinated patients was lower ( χ2=17.766, P<0.01) than those in non-liver injury group. The levels of platelet, hemoglobin, albumin and potassium in liver injury group were lower than those in non-liver injury group ( Z=-4.631, -2.368, -10.593 and -2.141, all P<0.05), while serum ALT, AST, γ-GT, urea nitrogen, glucose and hs-CRP levels were higher than those in the non-liver injury group ( Z=-7.451, -8.663, -4.410, -3.824, -3.278 and -3.884, all P<0.01). Multivariate Logistic regression analysis showed that age ( OR=2.580, 95% CI 1.429-4.657, P=0.002), history of diabetes ( OR=3.650, 95% CI 1.698-7.849, P=0.001), and decreased hemoglobin ( OR=1.993, 95% CI 1.066-3.726, P=0.031) and increased hs-CRP ( OR=1.797, 95% CI 1.283-2.517, P=0.001) were risk factors associated with liver function injury, while vaccination ( OR=0.499, 95% CI 0.312-0.798, P=0.004) was the protective factor for liver function. Conclusion:Liver function injury is frequently observed in COVID-19 patients infected with omicron variant, which is linked to age, underlying disease, and elevated inflammatory markers; while vaccination can lower the risk of liver injury in infected patients.